橙皮素对舌癌细胞的生长抑制作用及其机制

doi:10.13481/j.1671-587x.20150324

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (03): 558-562.doi: 10.13481/j.1671-587x.20150324

橙皮素对舌癌细胞的生长抑制作用及其机制

刘治慧¹, 杨巍^2,3

1. 台州学院附属市立医院口腔科, 浙江台州 318000;
2. 北华大学医学检验学院免疫学教研室, 吉林吉林 132013;
3. 台州学院医学院检验系, 浙江台州 318000

收稿日期:2014-12-01 发布日期:2015-08-01
通讯作者: 杨巍,副教授(Tel:0432-64608115,E-mail:yangwei660400@aliyun.com) E-mail:yangwei660400@aliyun.com
作者简介:刘治慧(1974-),男,浙江省台州市人,主任医师,副教授,主要从事口腔及颌面外科肿瘤治疗方面的研究。
基金资助:
浙江省中医药管理局中医药优秀青年人才基金资助课题(2013ZQ030);浙江省台州市科技局科技发展计划项目资助课题(131ky17)

Suppressive effect of hesperetin on growth of tongue cancer cells and its mechanism

LIU Zhihui¹, YANG Wei^2,3

1. Department of Stomatology, Affiliated Municipal Hospital, Taizhou University, Taizhou 318000, China;
2. Department of Immunology, School of Laboratoy Medicine, Beihua University, Jilin 132013, China;
3. Department of Laboratory Medicine, School of Medical Sciences, Taizhou University, Taizhou 318000, China

Received:2014-12-01 Published:2015-08-01

摘要/Abstract

摘要：

目的:检测橙皮素对舌癌细胞生长及Notch1活化的影响,探讨橙皮素抑癌作用机制。方法:将培养的Tca8113细胞分为DMSO对照组和25、50、75、100和125 μmol·L^-1橙皮素处理组。MTT法检测不同浓度橙皮素作用后Tca8113细胞增殖抑制率,流式细胞术检测各组细胞凋亡率和细胞周期分布,定量PCR和免疫印迹分别用于分析橙皮素作用前后Tca8113细胞中Notch1和Hes-1 mRNA及蛋白表达。结果:与DMSO对照组比较,125 μmol·L^-1橙皮素处理组24、48和72 h Tca8113细胞增殖抑制率明显升高(7.88%±1.90%、26.28%±2.2%和47.05%±1.90%)(P<0.05);在橙皮素处理72 h后,25、50、75、100 和125 μmol·L^-1橙皮素处理组细胞增殖抑制率分别为(6.14±1.20)%、(28.69±2.10)%、(28.33±2.60)%、(45.26±3.00)%和(47.05±1.90)%,橙皮素对Tca8113细胞的增殖抑制作用呈现时间剂量依赖性。流式细胞术,橙皮素作用72 h后, 0、50和100 μmol·L^-1橙皮素处理组细胞凋亡百分率从 2.9%±0.5%升高至23.4%±1.7%和 35.1%± 1.9%(P<0.05);G₀/G₁期细胞由(18.6±1.3)%升高至(33.4±1.5)%和(40.5±1.9)%(P<0.05), S期细胞百分率由(70.3±2.5)%下降至(56.8±2.0)%和(48.7±1.8)%(P<0.05)。定量PCR和免疫印迹分析,与DMSO对照组比较,橙皮素作用后Tca8113细胞中Notch1和Hes-1 mRNA及蛋白表达水平明显升高(P<0.05)。结论:橙皮素能够抑制Tca8113细胞增殖并诱导其发生细胞凋亡和细胞周期G₀/G₁期阻滞,其机制可能与活化Notch1有关联。

关键词: 橙皮素, 舌肿瘤, Notch1, Hes-1

Abstract:

Objective To detect the influence of hesperetin in the growth of tongue cancer cells and the activation of Notch1 signaling and to explore its mechanism. Methods The Tca8113 cells were cultured and divided into DMSO control and 25, 50, 75, 100, 125 μmol·L^-1 hesperetin groups.Then MTT assay was performed to detect the inhibitory rate of proliferation of Tca8113 cells after treatment with different concentrations of hesperetin.Flow cytometry was used to detect the the percentage of apoptotic cells and the distribution of cell cycle, and qPCR and Western blotting Methods were used respectively to analyze the relative expression levels of Notch1 and Hes-1 mRNA and protein. Results The MTT Results revealed that compared with DMSO control group, the inhibitory rates of proliferation of Tca8113 cells in 125 μmol·L^-1 hesperetin groups at 24, 48, and 72 h(7.88%±1.9%, 26.28%±2.2%, 47.05%±1.9%)were significantly increased(P<0.05).Moreover, after hesperetin treatment for 72 h, the inhibitory rates of proliferation of the Tca8113 cells in 25, 50, 75, 100, and 125 μmol·L^-1 hesperetin groups were (6.14±1.2)%, (28.69±2.1)%, (28.33±2.6)%, (45.26±3.0)%, and (47.05±1.9)%, respectively; hesperetin inhibited the growth of Tca8113 cells in a time- and dose-dependent manner.The flow cytometry Results showed that after hesperetin treatment for 72 h, compared with DMSO control group, the apoptotic rates of the Tca8113 cells in 50 and 100 μmol·L^-1 hesperetin groups were increased significantly from (2.9±0.5)% to (23.4±1.7)% and (35.1±1.9)%(P<0.05);the percentages of the Tca8113 cells in G₀/G₁ phage in 50 and 100 μmol·L^-1 hesperetin groups were increased significantly from (18.6±1.3)% to (33.4±1.5)% and (40.5±1.9)% (P<0.05), and the percentages of the Tca8113 cells in S phage were decreased from (70.3±2.5)% to (56.8±2.0)% and (48.7±1.8)%(P<0.05).The qPCR and Western blotting Results showed that the expression levels of Notch1 and Hes-1 mRNA and protein were all increased compared with DMSO control group(P<0.05). Conclusion Hesperetin can inhibit the growth and induce the apoptosis and G₀/G₁ arrest in the Tca8113 cells, and its mechanism may be related to activating the Notch1 signaling.

Key words: hesperetin, tongue neoplasms, Notch1, Hes-1

中图分类号:

R739.86

刘治慧, 杨巍. 橙皮素对舌癌细胞的生长抑制作用及其机制[J]. 吉林大学学报(医学版), 2015, 41(03): 558-562.

LIU Zhihui, YANG Wei. Suppressive effect of hesperetin on growth of tongue cancer cells and its mechanism[J]. Journal of Jilin University Medicine Edition, 2015, 41(03): 558-562.

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橙皮素对舌癌细胞的生长抑制作用及其机制

Suppressive effect of hesperetin on growth of tongue cancer cells and its mechanism

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